[Histonet] RE: IF on FFPE

[Preiszner, Johanna]

Hi Netters,

I've read the paper, but it's not clear what temperature they reached during the first round of HIER. They do not give the volume of the first HIER solution, only the time and wattage of the microwave treatment.
And which procedure do they call "first" and "second"? It looks different in the "Methods" section from what they mean in the "Discussion".

Hanna Preiszner


________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Martin, Erin [Erin.Martin <@t> ucsf.edu]
Sent: Tuesday, March 19, 2013 10:32 AM
To: histonet
Subject: [Histonet] IF on FFPE

Hi everyone, Several people have contacted me regarding the article I mentioned in my post yesterday.  Here are the details:



Immunofluorescence With Dual Microwave Retrieval of Paraffin-Embedded Sections in the Assessment of Human Renal Biopsy Specimens

AJCP 2013 139:71-78; doi:10.1309/AJCPRZG8EXN7BAID

Suozhu Shi, Qingli Cheng, Ping Zhang, Nan Wang, Ying Zheng, Xue-Yuan Bai, and Xiangmei Chen

Abstract: Immunofluorescence of frozen tissue sections (IF-F) is a classic technique for renal immunopathologic examination. However, it has certain disadvantages, such as diffuse antigen distribution and few or even no glomeruli in the section. We developed a new technique of immunofluorescence staining using dual microwave retrieval in paraffin-embedded renal tissue sections (IF-DMP) and compared IF-DMP with IF-F in 406 renal biopsy samples. IF-DMP detected significantly more glomeruli than did IF-F (P< .001). There was no significant difference for the specificity and sensitivity in the detection of immunoglobulins, complements, κ, and λ between IF-F and IF-DMP. Concordant observations were 98% for all immunofluorescence, complements, κ, and λ staining and 100% for immunoglobulin staining. Both techniques were completely accurate in confirming diagnoses of various glomerular diseases. IF-DMP provided clearer images of tissue structure and more precise localization of antigens, and it is a suitable alternative for traditional IF-F in clinical renal immunopathologic diagnosis.



This is all foreign to me - we do IF on derm following an inherited protocol.  I've never worked up any IF.  If anyone has thoughts on how to apply this to skin I'd appreciate it!



Thanks

Erin





Erin Martin, Histology Supervisor
UCSF  Dermatopathology Service
415-353-7248

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